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    Amperometric L-arginine biosensor based on a novel recombinant arginine deiminase


    Zhybak, Mykhailo T. and Fayura, Lyubov Y. and Boretsky, Yuriy R. and Gonchar, Mykhailo V. and Sibirny, Andriy A. and Dempsey, Eithne and Turner, Anthony P.F. and Korpan, Yaroslav I. (2017) Amperometric L-arginine biosensor based on a novel recombinant arginine deiminase. Microchimica Acta, 184 (8). pp. 2679-2686. ISSN 1436-5073

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    Abstract

    The authors describe an amperometric biosensor for the amino acid L-arginine (L-Arg). It is based on the use of a Nafion/Polyaniline (PANi) composite on a platinum screen-printed electrode (Pt-SPE) using a novel recombinant arginine deiminase isolated from Mycoplasma hominis. The protein was over-expressed, purified and employed as a biorecognition element of the sensor. Enzymatic hydrolysis of L-Arg leads to the formation of ammonium ions which diffuse into the Nafion/PANi layer and induce the electroreduction of PANi at a potential of -0.35 V (vs Ag/AgCl). L -Arg sensitivity was 684±32 A⋅M-1⋅m-2, and the apparent Michaelis-Menten constant (KMapp) 0.31±0.05 mM. The calibration plot was linear over the range 3 - 200 μM L-Arg, the limit of detection was 1 μM, and the response time (for 90% of the total signal change to occur) 15 s. The sensor was selective and exhibited good storage stability (> 1 month without loss in signal). The biosensor was applied to the analysis of L-Arg in pharmaceutical samples and of ammonium and L-Arg in spiked human plasma obtained from blood of healthy volunteers and those with a hepatic disorder. Data generated was found to be in good agreement with a reference fluorometric enzymatic assay.

    Item Type: Article
    Additional Information: This is the preprint version of the published article, which is available at: Zhybak, M.T., Fayura, L.Y., Boretsky, Y.R. et al. Microchim Acta (2017) 184: 2679. https://doi.org/10.1007/s00604-017-2290-4
    Keywords: L-Arginine; ammonium ions; amperometry; nafion; polyaniline; recombinantenzyme; plazma samples; pharmaceuticals;
    Academic Unit: Faculty of Science and Engineering > Chemistry
    Item ID: 11437
    Identification Number: https://doi.org/10.1007/s00604-017-2290-4
    Depositing User: Eithne Dempsey
    Date Deposited: 21 Oct 2019 17:04
    Journal or Publication Title: Microchimica Acta
    Publisher: Springer
    Refereed: Yes
    URI:

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